Effect of feeding different sources of rumen-protected methionine on milk production and N-utilization in lactating dairy cows.

Objectives of this study were to quantify production responses of lactating dairy cows to supplying absorbable Met as isopropyl-2-hydroxy-4-(methylthio)-butanoic acid (HMBi), or rumen-protected Met (RPM, Smartamine M; Adisseo, Alpharetta, GA) fed with or without 2-hydroxy-4-(methylthio)-butanoic acid (HMB), and to determine whether Met supplementation will allow the feeding of reduced dietary crude protein (CP). Seventy cows were blocked by parity and days in milk into 14 blocks and randomly assigned within blocks to 1 of the 5 dietary treatments based on alfalfa and corn silages plus high-moisture corn: 1 diet with 15.6% CP and no Met source (negative control); 3 diets with 15.6% CP plus 0.17% HMBi, 0.06% RPM + 0.10% HMB, or 0.06% RPM alone; and 1 diet with 16.8% CP and no Met supplement (positive control). Assuming that 50% of ingested HMBi was absorbed from the gastrointestinal tract and 80% of the Met in RPM was absorbed at intestine, the HMBi and RPM supplements increased metabolizable Met supply by 9 g/d and improved the Lys:Met ratio from 3.6 to 3.0. After a 2-wk covariate period during which all cows received the same diet, cows were fed test diets continuously for 12 wk. Diet did not affect dry matter intake (mean ± SD, 25.0±0.3 kg/d), body weight gain (0.59±0.2 kg/d), or milk yield (41.7±0.6 kg/d). However, feeding HMBi increased yield of energy-corrected milk and milk content of protein and solids-not-fat. Moreover, trends were observed for increased milk fat content and yield of fat and true protein on all 3 diets containing supplemental Met. Apparent N efficiency (milk N/N intake) was highest on the RPM treatment. Feeding 16.8% CP without a Met source elevated milk urea N and urinary excretion of urea N and total N and reduced apparent N efficiency from 34.5 to 30.2%, without improving production. Overall results suggested that feeding HMBi or RPM would give similar improvements in milk production and N utilization.

Effect on production of replacing dietary starch with sucrose in lactating dairy cows.

Replacing dietary starch with sugar has been reported to improve production in dairy cows. Two sets of 24 Holstein cows averaging 41 kg/d of milk were fed a covariate diet, blocked by days in milk, and randomly assigned in 2 phases to 4 groups of 6 cows each. Cows were fed experimental diets containing [dry matter (DM) basis]: 39% alfalfa silage, 21% corn silage, 21% rolled high-moisture shelled corn, 9% soybean meal, 2% fat, 1% vitamin-mineral supplement, 7.5% supplemental nonstructural carbohydrate, 16.7% crude protein, and 30% neutral detergent fiber. Nonstructural carbohydrates added to the 4 diets were 1) 7.5% corn starch, 0% sucrose; 2) 5.0% starch, 2.5% sucrose; 3) 2.5% starch, 5.0% sucrose; or 4) 0% starch, 7.5% sucrose. Cows were fed the experimental diets for 8 wk. There were linear increases in DM intake and milk fat content and yield, and linear decreases in ruminal concentrations of ammonia and branched-chain volatile fatty acids, and urinary excretion of urea-N and total N, and urinary urea-N as a proportion of total N, as sucrose replaced corn starch in the diet. Despite these changes, there was no effect of diet on microbial protein formation, estimated from total purine flow at the omasum or purine derivative excretion in the urine, and there were linear decreases in both milk/DM intake and milk N/N-intake when sucrose replaced dietary starch. However, expressing efficiency as fat-corrected milk/DM intake or solids-corrected milk/DM intake indicated that there was no effect of sucrose addition on nutrient utilization. Replacing dietary starch with sucrose increased fat secretion, apparently via increased energy supply because of greater intake. Positive responses normally correlated with improved ruminal N efficiency that were altered by sucrose feeding were not associated with increased protein secretion in this trial.

Milk and cheese from cows fed calcium salts of palm and fish oil alone or in combination with soybean products.

Twenty cows were used in a randomized block design experiment for 6 wk to determine the influence of feeding partial ruminally inert Ca salts of palm and fish oil (Ca-PFO), alone or in combination with extruded full-fat soybeans or soybean oil, on milk fatty acid (FA) methyl esters composition and consumer acceptability of milk and Cheddar cheese. Cows were fed either a diet containing 44% forage and 56% concentrate (control) or a diet supplemented with 2.7% Ca-PFO (FO), 5% extruded full-fat soybeans + 2.7% Ca-PFO (FOESM), or 0.75% soybean oil + 2.7% Ca-PFO (FOSO). Total dietary FA content in the control, FO, FOESM, and FOSO diets were 4.61, 6.28, 6.77, and 6.62 g/100 g, respectively. There was no difference in nutrient intake, milk yield, or milk composition among treatments. Conjugated linoleic acid (CLA) C(18:2) cis-9, trans-11 isomer, C(18:1) trans-11 (VA), and total n-3 FA in milk from cows on the control, FO, FOESM, and FOSO treatments were 0.56, 1.20, 1.36, and 1.74; 3.29, 4.66, 6.34, and 7.81; 0.62, 0.69, 0.69, and 0.67 g/100 g of FA, respectively. Concentrations of CLA, VA, and total n-3 FA in cheese were similar to milk. A trained sensory panel detected no difference in flavors of milk and cheese, except for acid flavor below a slightly perceptible level in cheese from all treatments. Results suggest that feeding Ca-PFO alone or in combination with extruded full-fat soybeans or soybean oil enhanced the CLA, VA, total unsaturated and n-3 FA in milk and cheese without negatively affecting cow performance and consumer acceptability characteristics of milk and cheese.

Comparison of calcium salts and formaldehyde-protected conjugated linoleic Acid in inducing milk fat depression.

Abomasal infusion studies have shown that trans-10, cis-12 conjugated linoleic acid (CLA) decreases milk fat synthesis. However, supplements of CLA must avoid rumen biohydrogenation for this technology to be applied to ruminants. Rumen protection methods would reduce CLA metabolism in the rumen and increase its supply to the small intestine. Our objective was to compare the efficacy of 2 forms of rumen-protected CLA at inducing milk fat depression. Three mid to late lactation Holstein cows each fitted with a rumen fistula were used in a 3 x 3 Latin square design. Treatments were: 1) control, 2) calcium salts of CLA (Ca-CLA), and 3) formaldehyde-protected CLA (FP-CLA). Supplements were designed to provide 10 g/d of trans-10, cis-12 CLA and were administered intraruminally once per day to ensure exact delivery of amount. Both CLA treatments substantially reduced milk fat yield and content compared with control, with the reductions in milk fat yield averaging 34% for the Ca-CLA treatment and 44% for the FP-CLA treatment. In contrast, milk yield, milk protein yield, and dry matter intake were unaltered by CLA treatment. Efficiency of transfer of trans-10, cis-12 CLA from the supplement into milk fat was 3.2 and 7.0% for Ca-CLA and FP-CLA, respectively. These values are much lower than transfer efficiencies reported for abomasally infused CLA, suggesting that much of the trans-10, cis-12 CLA present in the 2 formulations was biohydrogenated in the rumen. Overall, the extent of the reduction in milk fat yield indicates that both protection formulations are acceptable methods for the formulation of CLA supplements to induce milk fat depression in lactating dairy cows.

Effects of replacing dietary starch with sucrose on ruminal fermentation and nitrogen metabolism in continuous culture.

A dual-effluent continuous-culture system was used to evaluate the effects of partially replacing cornstarch with sucrose in a total mixed ration on ruminal fermentation and N metabolism. The 4 treatments were 0 (control), 2.5, 5.0, and 7.5% sucrose and, respectively, 7.5 (control), 5.0, 2.5, and 0% cornstarch in a total mixed ration containing 20% corn silage and 40% alfalfa silage. Fermenters were fed 4 times a day during four 9-d periods with sampling beginning on d 6. Replacing cornstarch with sucrose did not alter ruminal pH (5.97), total volatile fatty acids (VFA) (104.4 mmol/L), or the acetate to propionate ratio (2.16); however, branched-chain volatile fatty acids were higher for the control treatment compared with the 7.5% sucrose treatment. Five hours postfeeding, sucrose treatments significantly altered molar proportions of all volatile fatty acids, and acetate-to-propionate and glucogenic-to-lipogenic ratios. Digestibility of dry matter and N were not affected by treatment, but digestibility of total non-structural carbohydrates was increased with sucrose treatments. A quadratic effect was noted for neutral detergent fiber (NDF) digestibility as sucrose replaced starch. A higher NDF digestibility (66.1 vs. 59.9%) was observed for the 7.5% sucrose treatment compared with the other 2 sucrose treatments. Levels of ammonia N were within an acceptable range to support microbial protein synthesis and did not differ among treatments (mean=9.23 mg/dL). Sucrose inclusion in the total mixed ration did not affect bacterial N synthesis. Results indicate that (at the levels tested in this study) inclusion of sucrose in the diet when rumen-degradable protein is adequate does not affect ruminal fermentation.

Production and metabolic responses of periparturient Holstein cows to dietary conjugated linoleic acid and trans-octadecenoic acids.

Thirty-eight multiparous Holstein cows were utilized in a completely randomized design to examine the effect of feeding calcium salts of conjugated linoleic acid (CLA) and trans-octadecenoic acids (trans-C18:1) on animal performance and lipid and glucose metabolism during the transition to lactation. Dietary treatments were initiated approximately 28 d prior to expected calving dates and continued through d 49 postpartum. Prepartum treatments consisted of 1) a basal diet (Control), 2) basal diet + 150 g/d of CLA mix (CLA), and 3) basal diet + 150 g/d of trans-C18:1 mix (TRANS). Amounts of calcium salts of CLA and trans-C18:1 mixes were adjusted to 225 g/d during the 49-d postpartum treatment period. All diets were offered as a total mixed ration. Prepartum fat supplementation had no detectable effects on dry matter intake, body weight, or body condition score. After parturition, cows in the TRANS group consumed less dry matter at wk 4, 5, and 6 of lactation than did cows in the control group. Cows fed the trans-C18:1 supplement were in a more severe negative energy balance than those fed the control diet at 1 wk of lactation. Periparturient fat supplementation had no detectable effects on milk yield during wk 1 to 7 of lactation. Milk fat was not affected during wk 1 to 4, but was reduced after wk 4 of lactation by dietary CLA. Feeding calcium salts of CLA decreased short- to medium-chain fatty acid (C4 to C14) concentrations and increased both linoleic and linolenic acid concentrations in milk fat. Concentrations of nonesterified fatty acids and beta-hydroxybutyric acid in blood were greater in cows fed the CLA-supplemented diet than in those fed the control diet at 1 wk of lactation. In spite of small numerical tendencies, hepatic lipid and triacylglycerol concentrations did not vary significantly among dietary treatments. Periparturient fat supplementation had no detectable effects on plasma glucose and insulin concentrations. Steady-state concentrations of hepatic mRNA encoding pyruvate carboxylase and phosphoenolpyruvate carboxykinase were greater for the TRANS treatment group than the control and CLA groups. Results indicate that dietary CLA and trans-C18:1 fatty acids may affect lipid and glucose metabolism in early postpartum Holstein cows through distinct mechanisms.

In vitro determination of ruminal protein degradation using freeze-stored ruminal microorganisms.

Mixed ruminal microorganisms were harvested from a lactating dairy cow and preserved frozen or lyophilized. Fermentation characteristics of freshly strained ruminal fluid, frozen microorganisms, or lyophilized microorganisms were evaluated during a 24-h pre-incubation and a 4-h incubation with test proteins. Differences observed during the first 4 to 6 h in total amino acid concentration, optical density, pH and VFA concentrations, acetate:propionate ratio, and lactate concentration largely disappeared later in the pre-incubation. Protein degradation rates determined for expeller and solvent meals were .015 and .092 h-1, .015 and .101 h-1, and .005 and .019 h-1, with fresh ruminal fluid, frozen microorganisms, and lyophilized microorganisms, respectively. Regression of degradation rates obtained with fresh ruminal fluid on those obtained with pre-incubated, frozen microorganisms indicated the two methods were well correlated (r2 = .98 and .94 in two experiments). Mean in vitro degradability obtained for 17 feeds using pre-incubated, frozen microbes was 89% of that obtained using the in situ method; however, in situ degradation rates for these same feeds averaged only 67% of those obtained using frozen microorganisms. Ruminal undegraded protein values for nine samples of heated soybeans and soybean meal, determined using frozen microbes, were overestimated relative to in vivo values (in vivo = 1.1 + .8 in vitro; r2 = .77). These results indicated that ruminal microorganisms can be preserved by freezing and used as the inoculum for in vitro determination of ruminal protein degradation after overnight pre-incubation.

Preservation of ruminal microorganisms for in vitro determination of ruminal protein degradation.

Ruminal microorganisms, preserved either lyophilized or frozen, were compared with freshly strained ruminal fluid for proteolytic activity and as inoculum source for determination of ruminal protein degradation rates by the inhibitor in vitro method. Dialysis and glycerol addition had no effect on the proteolytic activity of preserved microorganisms. Net release of NH3 and total amino acids from protein using the fluid plus particle-associated microorganisms was higher than that found using the fluid-associated microorganisms alone. Method of inoculum preservation altered total proteolytic activity, but harvesting bacteria using centrifugal force greater than 5,000 x g did not increase proteolytic activity of the pellet. The proposed method for harvesting and preserving microorganisms consisted of centrifuging strained ruminal fluid at 5,000 x g (30 min at 4 degrees C), stirring the pellet in a 50:50 (vol/vol) solution of glycerol-McDougall's buffer for 15 min, and then storing at -20 degrees C. Protein degradation rates in incubations with preserved microorganisms were four to eight times slower than when using fresh ruminal fluid; however, feed proteins were ranked similarly for degradation rate. Preincubating the preserved microorganisms reduced blank concentrations of NH3 and total amino acid and increased protein degradative activity of the preserved inoculum. Degradation rates with preincubated, preserved inocula were similar to those obtained using fresh ruminal fluid. These results indicated that mixed ruminal microorganisms can be preserved by freezing and, after a preincubation period of 6 h, used as the inoculum source for in vitro estimation of ruminal protein degradation.

Characterization of the proteolytic activity of commercial proteases and strained ruminal fluid.

The objective of this research was to formulate a mixture of commercial proteases that would mimic the rate and extent of protein degradation obtained using strained ruminal fluid. The proteolytic activity of strained ruminal fluid and several commercial proteases was characterized using 13 L-amino acid p-nitroanilides as artificial substrates. A mixture of Streptomyces griseus protease, chymotrypsin, and proteinase K at .042, 2.5, and .5 enzyme units/mL, respectively, was similar to the activity of strained ruminal fluid against the same artificial substrates. However, degradative activities were different in incubations with feed proteins as substrates. The rates of degradation of expeller soybean meal, solvent soybean meal, and casein were .08, .05, and .08/h, respectively, using the enzyme mixture and .03, .15, and .24/h using strained ruminal fluid. A second experiment compared degradative activity of S. griseus protease at .066 enzyme units/mL, ficin at .5 enzyme units/mL, and a mixture of trypsin, carboxypeptidase B, chymotrypsin, and carboxypeptidase A at 116.6, .5, 2.5, and .5 enzyme units/mL, respectively. Protein degradation rates obtained with strained ruminal fluid were two to six times faster than those obtained with the enzyme mixtures. A third experiment compared the degradability of 15 feed proteins with the mixture of trypsin, carboxypeptidase B, chymotrypsin and carboxypeptidase A to that with strained ruminal fluid. Degradation rates obtained using strained ruminal fluid ranged from .007 to .217/h; degradation rates using the enzyme mixture ranged from .010 to .079/h and were lower (P = .004) than with strained ruminal fluid. Overall, the experiments indicated that the commercial enzymes tested did not mimic the protein degradative activity of strained ruminal fluid.

Preweaning intake and postweaning dietary energy effects on intake and metabolism of calves weaned at 26 days of age.

Holstein calves (n = 48) were used to evaluate whether preweaning feeding management affects postweaning DMI and metabolism after abrupt weaning at 26 d of age. Calves were fed only whole milk until d 19 of age. From d 19 to 26, calves were provided one of three isocaloric and isonitrogenous diets: milk at 15% of d-1 BW, milk at 12% of d-1 BW plus a grain-based supplement added to the pail, or milk at 12% of d-1 BW and drenched (force-fed) with the supplement. After weaning, calves were offered a low or high energy (1.7 or 5.8% ether extract) starter. From d 26 to 36, starter intake was similar between calves fed the supplement in the pail and those fed only milk but lower than for calves drenched with supplement. Plasma NEFA increased threefold from d 26 to 27, and calves fed only milk had higher NEFA than drenched calves. From d 26 to 42, plasma VFA of drenched calves were higher than those fed only milk. Except for an increase in plasma NEFA at d 36 and 42, high energy starter did not affect any variable. Preweaning drenching of calves stimulated postweaning adaptation to dry diets. Intake postweaning appears to depend more on physiological adaptation to dry diets than a preweaning feeding management.

Evaluation of starter diet crude protein level and feeding regimen for calves weaned at 26 days of age.

The objectives of this study were to evaluate two levels of CP in starter diets offered to calves weaned on d 26 and to compare postweaning intake of starter diets and performance of calves weaned at 26 or 42 d. Forty Holstein calves were housed in individual outdoor hutches and assigned to one of four treatments. Treatments consisted of weaning on d 26 after offering a 20% CP starter diet from d 1, or d 21, or after offering a 25% CP starter diet from d 21, or weaning on d 42 after offering a 20% CP starter diet from d 1 (control). Between d 42 and 84, calves were commingled in outdoor group pens. On d 42, BW of control calves was heavier than that of early weaned calves. On d 84, BW did not differ among treatments. Consumption of starter diet was not affected by the age at which it was first offered. Weaned calves consumed more starter diet than controls from d 29 to 42. Postweaning performance was not affected by increasing CP in the starter diet. Weaning treatment did not affect the incidence of scours. Comparison of CP intakes with current NRC recommendations suggests that CP requirements for young milk-fed calves are adequate.